Fig. 4
From: Regulation of glypican 6-mediated Wnt activation maintains TDP-43 nuclear localization in neurons
Gpc6 genetic reduction restores NCT and TDP-43 nuclear localization in Gde2KO animals. (a,b) Representative western blot images of GPC6 in Gpc6WT and Gpc6Het cortical extracts (a) and graph (b) quantifying GPC6 levels normalized to total protein. Arrow points to the band for furin cleaved GPC6. Gpc6WT vs. Gpc6Het: *p = 0.0215, n = 4, Unpaired t-test. (c,d). Representative images of immunostained cortical sections of 4-month-old WT;WNT-GFP-MYC, Gde2KO:WNT-GFP-MYC, and Gde2KO;Gpc6Het;WNT-GFP-MYC animals (c,c’) and graphs quantifying the proportion of MYC + neurons (d) and number of MYC + non-neuronal cells per field of view (e) in the cortex. Images in (c’) are magnified from (c). Scale bars: c = 50 μm, (c’) = 20 μm. (d) WT;WNT-GFP-MYC vs. Gde2KO:WNT-GFP-MYC: **p = 0.0044; Gde2KO:WNT-GFP-MYC vs. Gde2KO;Gpc6Het;WNT-GFP-MYC: **p = 0.0097; WT;WNT-GFP-MYC vs. Gde2KO;Gpc6Het;WNT-GFP-MYC: ns p = 0.7573. N = 4–5 animals, n ~ 600 neurons per animal. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. (e) WT;WNT-GFP-MYC vs. Gde2KO:WNT-GFP-MYC: ns p = 0.2294; Gde2KO:WNT-GFP-MYC vs. Gde2KO;Gpc6Het;WNT-GFP-MYC: ns p = 0.9888; WT;WNT-GFP-MYC vs. Gde2KO;Gpc6Het;WNT-GFP-MYC: ns p = 0.2794. N = 4–5 animals. (f, h) Representative images of immunostained cortical sections of 4-month-old WT, Gde2KO, and Gde2KO;Gpc6Het. Scale bar = 50 μm. Arrows highlight cells in Gde2KO exhibiting nuclear exclusion of Ran (f) and cytoplasmic mislocalization of TDP-43 (h). (g,i). Graph quantifying Ran N/C ratios (g. WT vs. Gde2KO: ***p = 0.0007; Gde2KO vs. Gde2KO;Gpc6Het: **p = 0.0039; WT vs. Gde2KO;Gpc6Het: ns p = 0.6670. Ordinary one-way ANOVA with Tukey’s multiple comparisons test) and TDP-43 N/C ratios (i. WT vs. Gde2KO: **p = 0.0070; Gde2KO vs. Gde2KO;Gpc6Het: *p = 0.0314; WT vs. Gde2KO;Gpc6Het: ns p = 0.9577. Brown-Forsythe and Welch ANOVA tests). N = 6 animals, ≥ 120 cells per animal. All graphs: mean ± sem.
